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基于金磁纳米粒子和石墨烯片的新型弓形虫特异性 IgM 免疫传感器的检测

A novel immunosensor for detecting toxoplasma gondii-specific IgM based on goldmag nanoparticles and graphene sheets.

机构信息

Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.

出版信息

Colloids Surf B Biointerfaces. 2013 Jan 1;101:481-6. doi: 10.1016/j.colsurfb.2012.07.021. Epub 2012 Jul 25.

Abstract

A novel electrochemical immunosensor for detecting toxoplasma gondii-specific IgM (Tg-IgM) was constructed based on goldmag (Au-Fe(3)O(4)) nanoparticles and graphene sheets (GS). Thionine (Thi), as a mediator, was first electropolymerized on a nafion-GS (Nf-GS) modified electrode. Subsequently, gold nanoparticles (AuNPs) were attached onto the poly-thionine film through π-stacking interactions, and then were used to immobilize toxoplasma gondii antigen (Tg-Ag) for immunosensor fabrication. A sandwich-type immunoassay for Tg-IgM was performed using Au-Fe(3)O(4) labeled anti-IgM-horseradish peroxidase (HRP) as trace label. Electrochemical detection was carried out in the presence of H(2)O(2) as HRP substrate. Using Au-Fe(3)O(4) provided a simple, non-chemical damaging method for regeneration, and enhanced the HRP reduction ability toward H(2)O(2). The AuNPs/Thi/Nf-GS nanocomposite also had good conductivity and biocompatibility, which effectively improved the immunosensor sensitivity. Under optimal conditions, the immunosensor can detect Tg-IgM in two linear ranges from 0.0375 to 1.2 AU mL(-1) and from 2.0 to 18 AU mL(-1) with a detection limit of 0.016 AU mL(-1) (S/N=3). The immunosensor exhibited good reproducibility, stability, and selectivity as well.

摘要

一种基于金磁(Au-Fe(3)O(4))纳米粒子和石墨烯片(GS)的检测刚地弓形虫特异性 IgM(Tg-IgM)的新型电化学免疫传感器构建完成。首先,在 Nafion-GS(Nf-GS)修饰电极上电化学聚合了硫堇(Thi)作为媒介。随后,通过π-堆积相互作用将金纳米粒子(AuNPs)附着在聚硫堇薄膜上,然后用于固定刚地弓形虫抗原(Tg-Ag)以制备免疫传感器。采用 Au-Fe(3)O(4)标记的抗 IgM-辣根过氧化物酶(HRP)作为示踪标记,进行 Tg-IgM 的三明治型免疫分析。在 H(2)O(2)作为 HRP 底物的存在下进行电化学检测。使用 Au-Fe(3)O(4)提供了一种简单、非化学破坏性的再生方法,并增强了 HRP 对 H(2)O(2)的还原能力。AuNPs/Thi/Nf-GS 纳米复合材料还具有良好的导电性和生物相容性,有效提高了免疫传感器的灵敏度。在最佳条件下,该免疫传感器可以在两个线性范围内检测 Tg-IgM,范围从 0.0375 到 1.2 AU mL(-1)和 2.0 到 18 AU mL(-1),检测限为 0.016 AU mL(-1)(S/N=3)。该免疫传感器还表现出良好的重现性、稳定性和选择性。

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